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Biomedical subjects

L Gern

Publications and source records attributed to L Gern.

At least 55 records · Page 3Linked to original sources
PubMed1995

Biology of Ixodes (Pholeoixodes) hexagonus under laboratory conditions. Part II. Effect of mating on feeding and fecundity of females.

The effect of mating on the feeding and fecundity of Ixodes (Pholeoixodes) hexagonus females was studied under controlled laboratory conditions of 22-23 degrees C and 98% relative humidity. The feeding period of mated females was 6-15 days and 11-13 days for unmated females. The mean weight of the engorged mated females was 114.84 +/- 45.89 mg, whereas, that of the engorged unmated females was significantly lower (80.61 +/- 28.84 mg). During the initial slow feeding period, the weight of mated females increased 6.6 times. At the end of the blood feeding, they had increased their initial weight 35.5 times. Unmated females never entered the rapid engorgement phase and up to 12 days of feeding period their mean weight did not increase more than 9.2 times. The pre-oviposition periods of mated and unmated females were 6-15 days and 4-12 days, respectively. The mean of the egg production efficiency was 40.26 +/- 12.47% for mated females and 35.68 +/- 12.2% for unmated females. The mean of the mass conversion efficiency was 73.6 +/- 13.7% for mated females and 66.48 +/- 16.55% for unmated females. Sixty per cent of the eggs deposited by mated females hatched whereas only 1% of the eggs deposited by unmated females hatched. These results indicate that I. hexagonus females possess some predisposition for parthenogenesis and only fertility and not fecundity depends on mating.

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PubMed1995

Strain variation of Lyme disease spirochetes isolated from Ixodes ricinus ticks and rodents collected in two endemic areas in Switzerland.

The relationship among Lyme borreliosis, Borrelia genospecies, rodent reservoirs, and Ixodes ricinus L. ticks was studied in two endemic areas in Switzerland. Ear punch biopsies and sampling of internal organs were used to isolate Borrelia burgdorferi (Johnson, Schmid, Hyde, Steigerwalt and Brenner) from small mammals, Apodemus sylvaticus L., A. flavicollis Melchoir, Clethrionomys glareolus Schreber. Spirochetes were isolated from ear tissue and spleen of the rodents. Isolates were homogeneous and belonged to typing group II identified as B. afzelii (Canica, Nato, du Merle, Mazie, Baranton and Postic). Our data show that a specific association exists between B. afzelii and rodent reservoirs in European foci. Borreliae were also isolated from field-collected I. ricinus ticks from the same study areas. Proteinic and antigenic analysis indicated that more than one genospecies were present in the tick population. This suggests that other vertebrate hosts may serve as reservoirs of other Borrelia genospecies implicated with Lyme disease.

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PubMed1994

Borrelia burgdorferi in rodents (Apodemus flavicollis and A. sylvaticus): duration and enhancement of infectivity for Ixodes ricinus ticks.

Ixodes ricinus is an important vector of Borrelia burgdorferi in Europe, and small rodents (Apodemus flavicollis, A. sylvaticus and Clethrionomys glareolus) are important sources for infecting ticks. In this study, we examined their reservoir role by studying the duration of their infectivity for ticks. A. flavicollis and A. sylvaticus mice captured in nature were exposed to uninfected I. ricinus larvae at different times after their capture: 10 days, and 2, 7, 11, 14 and 40 months. Ticks were examined for spirochaetes after moulting using direct immunofluorescence. All animals remained infective for ticks their life long but the efficiency of transmission from hosts to ticks varied from one individual to the other, presenting a three-fold variation (26.5% to 81.4%). Rodents continously exposed to successive infestations by larval I. ricinus ticks over a period of one month showed an enhancement of infectivity for larval ticks during this period.

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PubMed1994

Immunization of mice by recombinant OspA preparations and protection against Borrelia burgdorferi infection induced by Ixodes ricinus tick bites.

The wide distribution of Borrelia burgdorferi, the spirochete causing Lyme borreliosis, represents a human health hazard in many areas of the world. Vaccination has been proposed as an effective prevention strategy. Vaccination experiments were conducted with preparations of recombinant outer surface protein A (OspA) derived from Borrelia burgdorferi strain ZS7. Mice received three doses (1 microgram each) of the antigens adsorbed to aluminum hydroxide. A strong immune response to the vaccine antigen was observed. Mice were challenged after immunization, using Ixodes ricinus nymphal ticks infected with Borrelia burgdorferi strain ZS7. Infection was investigated by ear biopsy culture, xenodiagnosis with uninfected larvae and serological response to Borrelia burgdorferi antigens. All unimmunized control animals were found to be infected, while all immunized animals were found to be protected against infection by Borrelia burgdorferi. In addition, most adult ticks derived from nymphs that fed on immunized mice were found to be free of spirochetes.

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PubMed1994

Characterization of Borrelia burgdorferi isolated from different organs of Ixodes ricinus ticks collected in nature.

Borrelia burgdorferi was isolated from 22 out of 133 adult Ixodes ricinus ticks collected from vegetation at two sites in Switzerland. From 17 ticks, spirochetes could be isolated from more than one organ. When the different isolates obtained from one tick were compared by SDS-PAGE analysis, differences in the protein profiles were observed in 8 cases. The isolates were further compared by immunological methods using mono- and polyclonal antibodies. Differences were observed in the proteins of 31-35 kDa and 18-25 kDa. Genetic divergence among isolates was evaluated by use of a B. burgdorferi specific gene probe for ospA. Correlation could be observed between immunological differences in OspA defined by monoclonal antibody LA31 and genetic variation of ospA as judged by restriction fragment length polymorphism (RFLP). Our findings indicate that systemic infection in unfed I. ricinus adults, as reflected by isolation of B. burgdorferi from multiple organs of one tick, is more frequent (8/22, 36%) than previously described (5%). Moreover, the presence of different B. burgdorferi phenotypes/genotypes in one tick is described for the first time. The findings may have bearings (i) on the time of tick attachment required for spirochete transmission since borreliae are already present in the salivary glands of systemically infected ticks at the beginning of the blood meal and (ii) perhaps also on the diversity of B. burgdorferi phenotypes inoculated by these ticks.

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PubMed1994

Comparison in the immunological properties of Borrelia burgdorferi isolates from Ixodes ricinus derived from three endemic areas in Switzerland.

Borrelia burgdorferi isolates were obtained from Ixodes ricinus from three sites in Switzerland. They were examined by SDS-PAGE and immunoblotting. The phenotypes, in respect of three outer surface proteins (Osp), differed between the sites of collection. In site 1, most isolates had an OspA of 31 kDa and an OspB of 34 kDa; in site 2, isolates presenting an OspA of 33 kDa dominated and in site 3, the isolates with an OspA of 32 kDa and an OspB of 35 kDa were most frequent. This distribution differed significantly. About half of the isolates from sites 1 and 3 reacted with anti-OspA monoclonal antibody H5332 compared to 29% from site 2. Site 1 isolates reacted significantly more frequently (81%) with another anti-OspA monoclonal antibody LA-31 than isolates from site 3 (P < 0.0001). These findings have implications for the epidemiology of Lyme borreliosis, for the further development of serodiagnostic reagents and for the development of a vaccine.

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PubMed1994

Infection of field-collected Ixodes ricinus (Acari: Ixodidae) larvae with Borrelia burgdorferi in Switzerland.

Free-living larvae from natural areas in five regions in Switzerland were collected to determine the infection rate of Ixodes ricinus L. larvae by Borrelia burgdorferi Johnson. Larvae were examined for the spirochetes using direct fluorescent antibody assay. Of 652 larvae examined, spirochetes were detected in 20 (3.1%). No differences in infection rate among sites were detected. The low prevalence of B. burgdorferi in larvae, compared with higher infection rate in nymphs (12.8%) and adults (14.5%), suggests that transovarial transmission is inefficient. These results suggest that I. ricinus can serve as a reservoir for B. burgdorferi in nature. Further study is needed on the relative importance of ticks, compared with mammals and birds, as reservoirs for B. burgdorferi.

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PubMed1994

Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease.

Many isolates of Borrelia burgdorferi have been obtained from ticks and vertebrate tissues collected in North America and continental Europe but only one established culture of United Kingdom Borrelia burgdorferi has been recorded. In this paper we report the isolation of B. burgdorferi from one of 108 tick pools representing 733 ticks and eight-four tissue samples from twenty-six rodents collected in the U.K, and the subsequent failure to establish the isolate (from ticks collected in Fordingbridge) in culture. In contrast, using identical techniques and culture medium, B. burgdorferi was isolated from one of seven tick pools collected in Switzerland, and from a single pool of ticks collected in Slovakia, and both isolates were successfully passaged. Analysis of questing I. ricinus collected from Fordingbridge by direct immunofluorescence showed 6/32 (19%) of adults and 8/108 (7%) of nymphs were positive for B. burgdorferi, although only one nymph contained > or = 1000 spirochaetes. To examine further the problem of isolating U.K. B. burgdorferi, twelve Ixodes ricinus tick samples from Fordingbridge, a recognized focus of Lyme disease, were subjected to isolation and culturing techniques, and the procedures monitored by use of the polymerase chain reaction (PCR). Whereas 11/12 samples were PCR positive after 2 weeks in culture, only one was PCR positive after 4 weeks. Motile spirochaetes were not visible by dark-field microscopy in any of the cultures. The results indicate that the standard BSK II medium routinely used to isolate and culture B. burgdorferi does not readily support the replication of the Borrelia species endemic to the U.K.

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PubMed1993

Ability of transovarially and subsequent transstadially infected Ixodes hexagonus ticks to maintain and transmit Borrelia burgdorferi in the laboratory.

In a previous study, transstadial and transovarial survival of Borrelia burgdorferi in Ixodes hexagonus and transmission to laboratory mice via the bite of infected females were demonstrated. Here, we report the ability of I. hexagonus progeny infected transovarially to maintain and transmit the spirochaete to the host. Ticks were examined for spirochaetes by direct immunofluorescence antibody test. I. hexagonus larvae derived from the parental transstadially infected females were fed on two white mice: 21/54 (38.9%) of these ticks examined as unfed nymphs were infected. I. hexagonus nymphs were fed on three white mice and examined for spirochaetes after moulting as adults: 7/25 (28%) were found to harbour the spirochaete. The success of B. burgdorferi transmission to the mice by larval and nymphal I. hexagonus was determined by xenodiagnosis using I. ricinus larvae: 20/50 (40%) and 30/99 (30.3%) of the I. ricinus larvae fed on the mice infected by I. hexagonus larvae and nymphs respectively became infected. This study shows that B. burgdorferi can be maintained through transovarial and subsequent transstadial transmissions in I. hexagonus.

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PubMed1993

Distinct patterns of protective antibodies are generated against Borrelia burgdorferi in mice experimentally inoculated with high and low doses of antigen.

We have studied the development of clinical arthritis and the generation of protective antibodies in two normal, inbred strains of mice either infected by ticks or experimentally (subcutaneous) inoculated with increasing numbers of Borrelia burgdorferi organisms. AKR/N mice developed only mild and DBA/2 mice only marginal clinical arthritis irrespective of the route of infection or the numbers of spirochetes (10-10(8)) inoculated. In contrast, immunodeficient SCID mice developed severe chronic arthritis under similar conditions, but with a delayed onset at lower numbers of needle-inoculated spirochetes or after tick bite. AKR/N and DBA/2 mice inoculated with either 10(4) (and fewer) B. burgdorferi organisms or via experimentally infected ticks generated antibodies with specificities for a variety of B. burgdorferi antigens except those to the outer surface proteins A and B (OspA, OspB). In contrast, mice inoculated with more than 10(4) spirochetes (10(5)-10(8)) developed in addition antibodies to OspA and OspB. Most notably, all three types of immune sera taken from DBA/2 mice showed similar capacities to confer protection on SCID mice against subsequent challenge with viable B. burgdorferi organisms. The data not only demonstrate that the quality of humoral immune responses to B. burgdorferi in mice is determined by the antigenic load, they also indicate the existence of further protective antibodies with specificities distinct from OspA and OspB.

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PubMed1993

Immune sera to individual Borrelia burgdorferi isolates or recombinant OspA thereof protect SCID mice against infection with homologous strains but only partially or not at all against those of different OspA/OspB genotype.

The outer surface proteins OspA and OspB of Borrelia burgdorferi have recently been demonstrated to be major target proteins for protective antibodies in mice against infection with the homologous spirochaetal strain. However, it has become clear from a variety of studies that B. burgdorferi isolates of different geographical origin and/or sources are heterogeneous and that they can be divided into at least six subgroups according to their distinct OspA/OspB genotypes. In order to analyse cross-protection between these subgroups we have now generated immune sera to various isolates of B. burgdorferi with different OspA/OspB genotypes. We show that passive immunization with antisera specific for whole spirochaetes or recombinant OspA of one spirochaetal isolate protects severe combined immunodeficiency mice against infection with strains of the corresponding OspA/OspB genotype but only partially or not at all against infection with isolates expressing distinct OspA/OspB genotypes. The incomplete protection mediated by individual antisera against independent isolates of B. burgdorferi suggests that an effective subunit vaccine against Lyme disease should consist of a mixture of OspA structures covering the heterogeneity of this protein within the species B. burgdorferi.

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PubMed1993

Mode of inoculation of the Lyme disease agent Borrelia burgdorferi influences infection and immune responses in inbred strains of mice.

Mice were infected with Borrelia burgdorferi by infection via Ixodes ricinus and experimental inoculation to determine whether transmission rates of spirochetes and antibody responses are influenced. Mice infected by the natural route were substantially more infective for ticks; two- to sixfold more tick larvae were positive for B. burgdorferi than those fed on experimentally inoculated mice. In natural infection, spirochetemia may be greater or spirochetes may be more accessible for transmission. Thus, this form of xenodiagnosis could be used to determine levels of spirochetes in the vertebrate host. Similar levels of antibody were present in all mice; however, those infected by the natural route lacked antibodies to outer surface proteins (Osp) A and B. The small antigen dose given through a tick bite may not have been sufficient to induce rapid OspA or OspB antibodies, thereby allowing the later development of higher levels of spirochetemia.

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PubMed1993

Analysis of intra-specific variation in the fatty acid profiles of Borrelia burgdorferi.

Analysis of the fatty acid methyl esters (FAMEs) of bacteria is a commonly used chemotaxonomic technique. Application of this methodology to spirochaetes associated with Lyme borreliosis revealed distinct clusters corresponding to three genetically distinguished groups: Borrelia burgdorferi sensu stricto, B. garinii, and the VS461 group. However, B. garinii formed a common group with B. hermsii, a relapsing fever spirochaete, and VS461 grouped with B. turicatae and B. parkeri, two other relapsing fever spirochaetes. The diversity in fatty acid profiles of Lyme disease spirochaetes has implications for the protean clinical manifestations of the disease.

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PubMed1993

Ixodes ricinus immatures on birds in a focus of Lyme borreliosis.

The infestation of birds by immature Ixodes ricinus was studied during 6 months in a Swiss woodland, where Lyme borreliosis is endemic. Thirteen passerine species were found to be parasitized by I. ricinus subadults and specially Turdus merula, T. philomelos and Erithacus rubecula. Overall, 300 larvae and 162 nymphs were collected on 95 avian hosts. Prevalence of infestation of nymphs on birds was higher in spring; larvae peaked in summer. The infection of birds by Borrelia burgdorferi was also studied using blood cultivation and examination of ticks. Motionless spirochetes were isolated from two E. rubecula. Infected ticks were removed from five species of passerines, and mainly three species of Turdidae (T. merula, T. philomelos and E. rubecula). Infection rate of larvae and nymphs by spirochetes averaged 16.3% and 21.7%, respectively. These percentages, compared to the infection rate of questing ticks collected through dragging, suggest that some Turdidae may play a role as amplifying hosts for spirochetes in the focus.

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PubMed1993

Borrelia burgdorferi in a focus of Lyme borreliosis: epizootiologic contribution of small mammals.

The contribution of woodmice (Apodemus sylvaticus), yellow-necked mice (Apodemus flavicollis) and bank voles (Clethrionomys glareolus) was compared in a focus of Lyme borreliosis in Switzerland during a 7 months' study. All three species of mice and one species of shrews (Sorex araneus) were shown parasitized by infected Ixodes ricinus immatures. About 14% of larvae and 50% of nymphs collected on small mammals were infected with B. burgdorferi. Spirochetes were isolated from blood of 3 woodmice and one yellow-necked mouse. The infectious status of rodents was estimated by tick xenodiagnosis. Prevalence of infected rodents ranged from 20% to 44%. Mice presented a higher potential infectivity than voles. The prevalence of infected rodents showed a seasonal variation.

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PubMed1992

Changes in the protein profile and antigenicity of different Borrelia burgdorferi strains after reintroduction to Ixodes ricinus ticks.

Eight Swiss strains of Borrelia burgdorferi, with various protein profiles and the North-American strain B31 were artificially introduced into Ixodes ricinus ticks and reisolated 10 days later. All isolates were subsequently examined by SDS-PAGE analysis. Comparing initial isolates with the reisolates, we observed that 7 out of 9 strains changed their protein pattern with respect to the major proteins OspA, OspB and the 22 kDa protein after passage in the tick. The strains NE2, NE4 and NE83 with the initial phenotype of OspA and 22 kDa proteins changed to the phenotype of OspA and OspB, the strains B2 and NE202 with the initial phenotype of OspA acquired an additional protein of 22 kDa and the strain NE58 with the initial phenotype of OspA also acquired a protein of 22 kDa. Examination of these isolates by Western blot analysis demonstrated that the reaction with the monoclonal antibody H5332 and a monospecific polyclonal antibody PoAb/anti-22 kDa differed between the initial isolates and the reisolates.

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